1,25-(OH)_2D_3和全反式视黄酸联合应用对溃疡性结肠炎小鼠TREG细胞Foxp3表达的影响  

EFFECT OF 1,25-(OH)_2D_3 COMBINED WITH ALL-TRANS RETINOIC ACID ON FOXP3 EXPRESSION IN TREG CELLS IN MICE WITH ULCERATIVE COLITIS

作　　者：张小艳[1] 杨雯[1] 张英[1] 李秀花[1] 邱服斌[1] 

机构地区：[1]山西医科大学公共卫生学院,太原030001

出　　处：《营养学报》2016年第2期157-161,共5页Acta Nutrimenta Sinica

基　　金：国家自然科学基金项目(No.81573156);山西省基础研究计划项目(No.2014011044-1)

摘　　要：目的探讨1,25-(OH)2D3和全反式视黄酸(all-trans retinoic acid,ATRA)联合应用对溃疡性结肠炎小鼠Treg细胞Foxp3蛋白及其相关细胞因子的表达和血钙、肌酐水平的影响。方法 6w龄雌性C57BL/6小鼠50只,随机分为正常对照组(CN),溃疡性结肠炎模型组(model,M),1,25-(OH)_2D_3干预,全反式视黄酸(ATRA)干预,1,25-(OH)_2D_3+ATRA联合干预共5组,除CN组外,其余4组随意饮3%糖酐酯(DSS)溶液建立溃疡性结肠炎模型,从造模D3开始干预,干预剂1,25-(OH)_2D_3和ATRA分别用花生油溶解(后者需在暗光下进行),干预剂量1,25-(OH)_2D_3100ng(d/capita),ATRA 0.2 mg(d/capita),1,25-(OH)_2D_3+ATRA(100ng/d+0.2mg/d)每天对应灌胃干预,9d后处死小鼠,取血清和结肠组织。按小鼠一般情况进行疾病活动指数(disease activity index,DAI)评分;测量结肠长度;观察结肠大体形态和病理学损伤情况,并对其进行组织病理学评分;生物化学方法检测小鼠结肠髓过氧化物酶(myeloperoxidase,MPO)活性和血清钙、肌酐水平;免疫组化法测结肠组织Foxp3、TGF-β、IL-10蛋白表达情况。结果与M组比较,三个干预组小鼠溃疡性结肠炎大体形态改善,组织学损伤较轻,结肠长度缩短有所缓解,结肠组织MPO活性下降,Foxp3蛋白及其相关细胞因子TGF-β、IL-10表达水平上升,血钙和肌酐水平下降。且联合干预组相比单独干预组效果更明显,差异具有统计学意义(除血钙)。结论 1,25-(OH)_2D_3和全反式视黄酸(ATRA)联合应用能够增强Treg细胞Foxp3的表达,使Treg细胞更好地发挥抑制免疫应答的作用,并可能缓解1,25-(OH)_2D_3引起的血钙升高,改善肾功能。Objective To explore the effect of the combination of 1,25-（OH）_2D_3 and ATRA on the expression of Foxp3 in Treg cells in mice with ulcerative colitis induced by dextran sulfate sodium（DSS）. Methods Fifty 6-week-old C57BL/6 female mice were randomly divided into five groups： normal control group（CN）, ulcerative colitis model group（M）,1,25-（OH）_2D_3 group, ATRA group, 1,25-（OH）_2D_3 and ATRA group. Except CN,mice in the other four groups were all treated with DSS to induce ulcerative colitis and received their treatment daily by gavage. After nine days, all the mice were sacrificed. The serum and colon tissues were collected. The disease activity index（DAI）was evaluated according to the general situation. The colon length were measured. Macroscopic and histological score of colon were evaluated by observing the gross morphology and pathological injury of colon. The colon MPO activity and serum calcium,creatine level were measured by biochemical methods. The expression of Foxp3、TGF-β、IL-10 in colonic tissues were measured by immunohistochemistry. Results Compared with M, the DAI, gross and histological score and MPO activity were significantly lower in the other three treatment groups, and similarly, the colon length were longer,the expression of Foxp3,TGF-β,IL-10 protein were up-regulated in colonic tissues. The calcium and creatine levels in serum were reduced,but more effective action was observed the combination group,and the difference was statistically significant.（except for the calcium level）. Conclusion The combination of 1,25-（OH）_2D_3 and ATRA enhanced the protective effects on ulcerative colitis by up-regulating the Foxp3 expression in Treg cells, thus enhancing their suppressive effects on immune system. Possibly, the increase of serum calcium levels caused by 1,25-（OH）_2D_3 was reduced at the same time.

关 键 词：1 25-(OH)2D3 ATRA 溃疡性结肠炎 FOXP3 TGF-β IL-10 血钙 肌酐 

分 类 号：R574.62[医药卫生—消化系统]